Support is requested to continue the development and application of immobilized enzyme technology to problems in biochemical analysis and medical therapy. The concept of immobilized enzymes has been broadened in the investigator's laboratory to include the study of immobilized antibody and immobilized inorganic catalyst activity. The enzyme electrode project will exploit our ability to covalently bond enzyme to Teflon membrane etched by a suspension of sodium metal in naphthalene. A faciliated RIA system that eliminates all pipetting and centrifugation steps will be evaluated. We also will evaluate a 100-sample, microcomputer-controlled, completely automated system of RIA based on re-using gel-immobilized antibody. Evaluation of immobilized enzyme therapy using RBC-entrapped asparaginase to our laboratory has shown more than a two-fold prolongation of asparagine substrate suppression. These studies done in the monkey have shown good survival of the resealed RBC ghosts with a substantially greated T 1/2 for the immobilized blood enzyme activity. We now turn our attention to the immunological aspects of this type of immobilized enzyme therapy. Using a classical passive-immunity model of anaphylactic shock in the guinea pig, we plan to compare intravenous injection of asparaginase in solution form with the same amount of asparaginase injected entrapped in resealed autologous RBC ghosts. In the area of immobilized inorganic catalyst activity, we plan further development and evaluation of hemodialysis-based system of extracorporeal respiratory gas exchange based on oxygenation by transmembrane catalysis of hydrogen peroxide. BIBLIOGRAPHIC REFERENCES: Updike, S.J., Wakamiya, R.T., Lightfoot, E.N.: Action and survival of RBC-entrapped asparaginase in monkey. Science 193: 681-683, August 20, 1976. Updike, S.J., Chen, Y.-D. Mark, Shults, M.C.: Membrane oxygenation based on Transmembrane Catalysis of Hydrogen Peroxide. To be published in A.S.A.I.O. Vol. 23, 1977.